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Coding

Part:BBa_K782005

Designed by: Fedja Pavlovec   Group: iGEM12_Slovenia   (2012-09-19)

TALD:NLS DNA binding domain


  • TALD label represents TAL effector 1295 from zebrafish experiments (Sander et al., 2011).


Introduction

TAL effectors (TALEs) are bacterial plant pathogen transcription factors, that bind to DNA by specifically recognizing one base pair with a single tandem repeat in their DNA-binding domain. A tandem TALE repeat contains 33 to 35 amino acids, where the 12th and 13th amino acid, called a “repeat variable diresidue” (RVD), are responsible for specific interactions with the corresponding base pair (Scholze and Boch, 2011).

Svn 12 TALD--.png

Figure 1: Schematic representation of the construct.

Single binding sequence for TALD: tCGTCCAATAGCTTCTC


Characterization

TAL effector was provided by host lab and cloned into a biobrick vector. This construct was later used for designing TAL-based activator and repressor by adding VP16 and KRAB domain.


References

Sander, J. D., Cade, L., Khayter, C., Reyon, D., Peterson, R. T., Joung, J. K., and Yeh, J.-R. J. (2011) Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Nature Biotechnology 29, 697–698

Scholze, H., and Boch, J. (2011) TAL effectors are remote controls for gene activation. Curr. Opin. Microbiol. 14, 47-53


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2337
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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